P
US7172893B2ExpiredUtilityPatentIndex 98

Virus vectors and methods of making and administering the same

Assignee: UNIV NORTH CAROLINAPriority: Nov 10, 1998Filed: Jul 26, 2002Granted: Feb 6, 2007
Est. expiryNov 10, 2018(expired)· nominal 20-yr term from priority
Inventors:RABINOWITZ JOSEPH ESAMULSKI RICHARD JUDEXIAO WEIDONG
A61P 7/04A61P 37/04A61P 31/18A61P 7/00A61P 3/10A61P 35/00A61P 25/08A61P 25/00A61P 3/00A61P 25/28A61P 25/16A61P 25/14A61P 27/02C12N 2750/14145C12N 2810/60C12N 15/86A61P 11/00A61P 1/18Y10S977/804A61K 2039/525A61P 21/02C12N 2810/854C12N 2810/405C12N 2750/14143A61K 48/00C12N 2750/14245A61P 21/04C12N 2810/40C12N 2750/14043C12N 2750/14243C12N 2750/14222C12N 2750/14122A61K 39/00Y02A50/30
98
PatentIndex Score
328
Cited by
117
References
54
Claims

Abstract

The present invention provides genetically-engineered parvovirus capsids and viruses designed to introduce a heterologous gene into a target cell. The parvoviruses of the invention provide a repertoire of vectors with altered antigenic properties, packaging capabilities, and/or cellular tropisms as compared with current AAV vectors.

Claims

exact text as granted — not AI-modified
1. A hybrid virus particle comprising:
 a parvovirus capsid; and 
 a nucleic acid comprising at least one adeno-associated virus (AAV) serotype 2 inverted terminal repeat packaged within said parvovirus capsid, subject to the proviso that if said parvovirus capsid is an AAV capsid, the serotypes of said AAV capsid and said at least one AAV inverted terminal repeat are different. 
 
     
     
       2. The hybrid virus particle of  claim 1 , wherein said nucleic acid comprises at least one heterologous nucleotide sequence. 
     
     
       3. The hybrid virus particle of  claim 2  comprising two AAV inverted terminal repeats that flank said at least one heterologous nucleotide sequence. 
     
     
       4. The hybrid virus particle of  claim 2 , wherein said at least one heterologous nucleotide sequence encodes a protein or peptide. 
     
     
       5. The hybrid virus particle of  claim 4 , wherein said protein or peptide is a therapeutic protein or peptide. 
     
     
       6. The hybrid virus particle of  claim 4 , wherein said protein or peptide is an immunogenic protein or peptide. 
     
     
       7. The hybrid virus particle of  claim 4 , wherein said at least one heterologous nucleotide sequence encodes dystrophin, a mini-dystrophin, a clotting factor, β-glucocerebrosidase, erythropoietin, cystic fibrosis transmembrane regulator protein, a cytokine, β-globin, a hormone, α-globin or a growth factor. 
     
     
       8. The hybrid virus particle of  claim 2 , wherein said at least one heterologous nucleotide sequence encodes an untranslated RNA. 
     
     
       9. The hybrid virus particle of  claim 1 , wherein said parvovirus capsid is an autonomous parvovirus capsid. 
     
     
       10. The hybrid virus particle of  claim 1 , wherein said parvovirus capsid is a B19 capsid. 
     
     
       11. The hybrid virus particle of  claim 1 , wherein said parvovirus capsid is an AAV capsid. 
     
     
       12. The hybrid virus particle of  claim 11 , wherein:
 said AAV capsid is of a serotype selected from the group consisting of AAV serotypes 1, 3, 4, 5 and 6. 
 
     
     
       13. The hybrid virus particle of  claim 12  selected from the group consisting of:
 (a) a hybrid virus particle comprising an AAV serotype-1 capsid and at least one AAV serotype-2 inverted terminal repeat. 
 (b) a hybrid virus particle comprising an AAV serotype-3 capsid and at least one AAV serotype-2 inverted terminal repeat, 
 (c) a hybrid virus particle comprising an AAV serotype-4 capsid and at least one AAV serotype-2 inverted terminal repeat, 
 (d) a hybrid virus particle comprising an AAV serotype-5 capsid and at least one AAV serotype-2 inverted terminal repeat, and 
 (e) a hybrid virus particle comprising an AAV serotype-6 capsid and at least one AAV serotype-2 inverted terminal repeat. 
 
     
     
       14. The hybrid virus particle of  claim 1 , wherein said nucleic acid does not comprise AAV cap genes or AAV rep genes. 
     
     
       15. A pharmaceutical formulation comprising the hybrid virus particle of  claim 1  in a pharmaceutically-acceptable carrier. 
     
     
       16. An isolated nucleic acid for producing the hybrid virus particle of  claim 1 , wherein said isolated nucleic acid comprises parvovirus cap genes and adeno-associated virus (AAV) rep genes, subject to the proviso that if said parvovirus cap genes are AAV cap genes, the serotypes of said AAV cap genes and said AAV rep genes are different. 
     
     
       17. The isolated nucleic acid of  claim 16 , wherein said parvovirus cap genes are operably associated with an authentic parvovirus promoter. 
     
     
       18. The isolated nucleic acid of  claim 16 , wherein said parvovirus cap genes are B19 cap genes. 
     
     
       19. The isolated nucleic acid of  claim 18 , wherein said AAV rep genes are AAV serotype-2 rep genes. 
     
     
       20. The isolated nucleic acid of  claim 16 , wherein said cap genes are AAV cap genes. 
     
     
       21. The isolated nucleic acid of  claim 20 , wherein said AAV cap genes are operably associated with an authentic AAV promoter. 
     
     
       22. The isolated nucleic acid of  claim 21 , wherein said authentic AAV promoter is an AAV p40 promoter. 
     
     
       23. The isolated nucleic acid of  claim 20 , wherein:
 said AAV cap genes are of a serotype selected from the group consisting of AAV serotypes 1, 3, 4, 5 and 6; and 
 said AAV rep genes are of a serotype selected from the group consisting of AAV serotypes 1, 2, 3, 4, 5 and 6. 
 
     
     
       24. The isolated nucleic acid of  claim 23  selected from the group consisting of:
 (a) an isolated nucleic acid comprising AAV serotype-1 cap genes and AAV serotype-2 rep genes, 
 (b) an isolated nucleic acid comprising AAV serotype-3 cap genes and AAV serotype-2 rep genes, 
 (c) an isolated nucleic acid comprising AAV serotype-4 cap genes and AAV serotype-2 rep genes, 
 (d) an isolated nucleic acid comprising AAV serotype-5 cap genes and AAV serotype-2 rep genes, and 
 (e) an isolated nucleic acid comprising AAV serotype-6 cap genes and AAV serotype-2 rep genes. 
 
     
     
       25. A vector comprising the isolated nucleic acid of  claim 16 . 
     
     
       26. The vector of  claim 25 , wherein said vector is selected from the group consisting of plasmids, naked DNA vectors, bacterial artificial chromosomes, yeast artificial chromosomes, and viral vectors. 
     
     
       27. The vector of  claim 26 , wherein said vector is a plasmid. 
     
     
       28. A cell comprising the vector of  claim 25 . 
     
     
       29. The cell of  claim 28 , wherein said cell is selected from the group consisting of bacterial, protozoan, yeast, fungus, plant, and animal cells. 
     
     
       30. A method of delivering a nucleotide sequence to a cell, in vitro comprising introducing into a cell the hybrid virus particle according to  claim 2 . 
     
     
       31. The method of  claim 30 , wherein the heterologous nucleotide sequence is expressed in the cell. 
     
     
       32. The method of  claim 31 , wherein the protein or peptide is an immunogenic protein or peptide. 
     
     
       33. The method of  claim 30 , wherein the parvovirus capsid is a B19 capsid. 
     
     
       34. The method of  claim 30 , wherein the at least one heterologous nucleotide sequence encodes a protein or peptide. 
     
     
       35. The method of  claim 34 , wherein the protein or peptide is a therapeutic protein or peptide. 
     
     
       36. The method of  claim 34 , wherein the at least one heterologous nucleotide sequence encodes dystrophin, a mini-dystrophin, a clotting factor, β-glucocerebrosidase, or a growth factor. 
     
     
       37. The method of  claim 30 , wherein the heterologous nucleotide sequence encodes an untranslated RNA. 
     
     
       38. The method of  claim 30 , wherein the cell is selected from the group consisting of a neural cell, lung cell, retinal cell, epithelial cell, muscle cell, pancreatic cell, hepatic cell, myocardial cell, bone cell, spleen cell, keratinocyte, fibroblast, endothelial cell, prostate cell, germ cell, progenitor cell, and a stem cell. 
     
     
       39. The method of  claim 30 , wherein the parvovirus capsid is an AAV capsid. 
     
     
       40. The method of  claim 39 , wherein:
 the AAV capsid is of a serotype selected from the group consisting of AAV serotypes 1, 3, 4, 5 and 6. 
 
     
     
       41. The method of  claim 40 , wherein the hybrid virus particle is selected from the group consisting of:
 (a) a hybrid virus particle comprising an AAV serotype-1 capsid and at least one AAV serotype-2 inverted terminal repeat, 
 (b) a hybrid virus particle comprising an AAV serotype-3 capsid and at least one AAV serotype-2 inverted terminal repeat, 
 (c) a hybrid virus particle comprising an AAV serotype-4 capsid and at least one AAV serotype-2 inverted terminal repeat, 
 (d) a hybrid virus particle comprising an AAV serotype-5 capsid and at least one AAV serotype-2 inverted terminal repeat, 
 (e) a hybrid virus particle comprising an AAV serotype-6 capsid and at least one AAV serotype-2 inverted terminal repeat. 
 
     
     
       42. A cell comprising a vector comprising:
 parvovirus cap genes, 
 adeno-associated virus (AAV) rep genes, and 
 a nucleic acid comprising at least one AAV serotype 2 inverted terminal repeat, 
 subject to the proviso that if said parvovirus cap genes are AAV cap genes, said at least one AAV inverted terminal repeat is of a different AAV serotype than said cap genes. 
 
     
     
       43. The cell of  claim 42 , wherein said cell is a mammalian cell. 
     
     
       44. A cell comprising parvovirus cap genes and adeno-associated virus (AAV) rep genes stably integrated into the genome of the cell, subject to the proviso that if said parvovirus cap genes are AAV cap genes, the serotypes of said AAV cap genes and said AAV rep genes are different. 
     
     
       45. The cell of  claim 44  further comprising a nucleic acid comprising at least one AAV serotype 2 inverted terminal repeat, subject to the proviso that if said parvovirus cap genes are AAV cap genes, the serotypes of said AAV cap genes and said at least one AAV inverted terminal repeat are different. 
     
     
       46. A method of producing a hybrid virus particle, comprising:
 providing a cell with adeno-associated virus (AAV) rep genes, parvovirus cap genes, a nucleic acid comprising at least one AAV serotype 2 inverted terminal repeat, and helper functions for generating a productive AAV infection; subject to the proviso that if the parvovirus cap genes are AAV cap genes, the serotypes of the AAV cap genes and the at least one AAV inverted terminal repeat are different, and 
 allowing assembly of the hybrid virus particles. 
 
     
     
       47. The method of  claim 46 , further comprising collecting the hybrid virus particles. 
     
     
       48. The method of  claim 46 , wherein the nucleic acid comprises at least one heterologous nucleotide sequence. 
     
     
       49. The method of  claim 46 , wherein the parvovirus cap genes and AAV rep genes are provided by one or more transcomplementing packaging vectors. 
     
     
       50. The method of  claim 46 , wherein the parvovirus cap genes and AAV rep genes are provided by a plasmid. 
     
     
       51. The method of  claim 46 , wherein the parvovirus cap genes and AAV rep genes are provided by an adenovirus vector. 
     
     
       52. The method of  claim 46 , wherein the parvovirus cap genes and AAV rep genes are stably integrated into the genome of the cell. 
     
     
       53. The method of  claim 46 , wherein the parvovirus cap genes are AAV cap genes. 
     
     
       54. A hybrid virus particle produced by the method of  claim 46 .

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